AuthorsMiskin I, Rhodes G, Lawlor K, Saunders JR, Pickup RW
JournalMicrobiology
PubMed ID9782490
Prokaryote communities in post-glacial profundal freshwater sediments of Windermere, representing 10-12,000 years of deposition, were examined for culturability, viability and community structure. The potential for active geochemical cycles was inferred from the presence of specific groups of bacteria. Direct count procedures revealed 10(12) cells (g dry wt sediment)-1 in the ... More
5-Carboxyfluorescein diacetate as a probe for measuring the growth of keratinocytes.
AuthorsHanthamrongwit M, Reid WH, Courtney JM, Grant MH
JournalHum Exp Toxicol
PubMed ID8086226
There is a requirement for a convenient and reliable method for evaluating the growth rate of human keratinocytes cultured on collagen-based substrates. Therefore, three methods of determining cell growth were first used to quantify the growth rate of the well-characterised L929 mouse fibroblast cell line on tissue culture plastic and ... More
Fluorescent indicators of ion concentrations.
AuthorsTsien RY
JournalMethods Cell Biol
PubMed ID2538708
Rapid assessment of physiological status in Escherichia coli using fluorescent probes.
AuthorsPorter J, Edwards C, Pickup RW
JournalJ Appl Bacteriol
PubMed ID7592133
'Rapid and direct viability assessment of Escherichia coli in filtered, sterile lake water was possible using multiparameter flow cytometry. Fluorescent dyes were used as probes for different cellular functions (membrane potential, membrane integrity and intracellular enzyme activity), which were correlated with the ability of the cells to respond to nutrient ... More
Intercellular adhesion mediated by human muscle neural cell adhesion molecule: effects of alternative exon use.
'Mouse 3T3 fibroblasts were permanently transfected with cDNAs encoding isoforms of the neural cell adhesion molecule (N-CAM) present in human skeletal muscle and brain. Parental and transfected cells were then used in a range of adhesion assays. In the absence of external shear forces, transfection with cDNAs encoding either transmembrane ... More
Scavenger receptor BI mediates the selective uptake of oxidized cholesterol esters by rat liver.
AuthorsFluiter K, Sattler W, De Beer MC, Connell PM, van der Westhuyzen DR, van Berkel TJ
JournalJ Biol Chem
PubMed ID10085133
'High density lipoprotein (HDL) can protect low density lipoprotein (LDL) against oxidation. Oxidized cholesterol esters from LDL can be transferred to HDL and efficiently and selectively removed from the blood circulation by the liver and adrenal in vivo. In the present study, we investigated whether scavenger receptor BI (SR-BI) is ... More
Anatomical and physiological localization of prelabeled grafts in rat hippocampus.
AuthorsPyapali GK, Turner DA, Madison RD
JournalExp Neurol
PubMed ID1577121
'Dissociated rat fetal hippocampal cells were grafted into normal adult rats. The fetal cells were incubated with one of a number of fluorescent compounds at the time of the dissociation to facilitate identification of the individual grafted cells. The fluorescent labels which were analyzed for this purpose included rhodamine latex ... More
Pep7p provides a novel protein that functions in vesicle-mediated transport between the yeast Golgi and endosome.
AuthorsWebb GC, Zhang J, Garlow SJ, Wesp A, Riezman H, Jones EW
JournalMol Biol Cell
PubMed ID9168472
'Saccharomyces cerevisiae pep7 mutants are defective in transport of soluble vacuolar hydrolases to the lysosome-like vacuole. PEP7 is a nonessential gene that encodes a hydrophilic protein of 515 amino acids. A cysteine-rich tripartite motif in the N-terminal half of the polypeptide shows striking similarity to sequences found in many other ... More
Transfersomes--a novel vesicular carrier for enhanced transdermal delivery: development, characterization, and performance evaluation.
AuthorsJain S, Jain P, Umamaheshwari RB, Jain NK
JournalDrug Dev Ind Pharm
PubMed ID14606665
'This work describes the use of a novel vesicular drug carrier system called transfersomes, which is composed of phospholipid, surfactant, and water for enhanced transdermal delivery. The transfersomal system was much more efficient at delivering a low and high molecular weight drug to the skin in terms of quantity and ... More
Continuous measurement of mitochondrial pH gradients in isolated hepatocytes by difference ratio spectroscopy.
AuthorsThomas PJ, Gaspers LD, Pharr C, Thomas JA
JournalArch Biochem Biophys
PubMed ID1898020
'The pH gradient, delta pH, present across the inner mitochondrial membrane in isolated rat hepatocytes was continuously monitored with a novel spectroscopic technique that utilizes the weak acid fluorescein. Unlike most cytosolic pH indicators, such as 2'',7''-bis(carboxyethyl)-5,(6)-carboxyfluorescein (BCECF), fluorescein freely distributes between the cytosolic and mitochondrial compartments. As is typical ... More
A comparative study of carboxyfluorescein diacetate and carboxyfluorescein diacetate succinimidyl ester as indicators of bacterial activity.
AuthorsHoefel D, Grooby WL, Monis PT, Andrews S, Saint CP
JournalJ Microbiol Methods
PubMed ID12531507
'Staining bacteria with esterified fluorogenic substrates followed by flow cytometric analysis offers a means for rapid detection of metabolically active bacteria. Flow cytometry (FCM) was used to assess carboxyfluorescein diacetate (CFDA) and carboxyfluorescein diacetate succinimidyl ester (CFDA/SE) as indicators of bacterial activity for cultured bacteria, including Aeromonas hydrophila, Bacillus subtilis, ... More
Flow cytometric assessment of viability of lactic acid bacteria.
AuthorsBunthof CJ, Bloemen K, Breeuwer P, Rombouts FM, Abee T
JournalAppl Environ Microbiol
PubMed ID11319119
'The viability of lactic acid bacteria is crucial for their applications as dairy starters and as probiotics. We investigated the usefulness of flow cytometry (FCM) for viability assessment of lactic acid bacteria. The esterase substrate carboxyfluorescein diacetate (cFDA) and the dye exclusion DNA binding probes propidium iodide (PI) and TOTO-1 ... More
Confocal imaging of organic anion transport in intact rat choroid plexus.
AuthorsBreen CM, Sykes DB, Fricker G, Miller DS
JournalAm J Physiol Renal Physiol
PubMed ID11934698
'We used confocal microscopy and quantitative image analysis to follow the movement of the fluorescent organic anion fluorescein (FL) from bath to cell and cell to blood vessel in intact rat lateral choroid plexus. FL accumulation in epithelial cells and underlying vessels was rapid, concentrative, and reduced by other organic ... More
Movements of fluorescent probes in the mechanism of cell fusion induced by poly(ethylene glycol).
AuthorsAhkong QF, Desmazes JP, Georgescauld D, Lucy JA
JournalJ Cell Sci
PubMed ID3448101
'It has been claimed that purified poly(ethylene glycol) (PEG) is able only to aggregate cells and not to fuse them. In our hands, purified PEG 6000 (recrystallized/dialysed) induces both aggregation and fusion of human erythrocytes, and the mechanism of fusion by the purified polymer has been investigated with fluorescent probes. ... More
Structural organization of interphase 3T3 fibroblasts studied by total internal reflection fluorescence microscopy.
AuthorsLanni F, Waggoner AS, Taylor DL
JournalJ Cell Biol
PubMed ID3980580
'We studied the laminar organization of 3T3 fibroblast cells growing on glass slides by use of total internal reflection illumination to excite fluorescence emission (TIRF) from labeled molecules and stained cellular compartments that are very close to the cell-substrate contact region. Mitochondria, distant from the contact regions and stained with ... More
Real-time multi-wavelength fluorescence imaging of living cells.
AuthorsMorris SJ
JournalBiotechniques
PubMed ID2331370
'We describe a new real-time fluorescence video microscope design for capturing intensified images of cells containing dual wavelength "ratio" dyes or multiple dyes. The microscope will perform real-time capture of two separate fluorescence emission images simultaneously, improving the time resolution of spatial distribution of fluorescence to video frame rates (30 ... More
Millimeter wave exposure reverses TPA suppression of gap junction intercellular communication in HaCaT human keratinocytes.
AuthorsChen Q, Zeng QL, Lu DQ, Chiang H
JournalBioelectromagnetics
PubMed ID14696047
'The effect of 30.16 GHz millimeter wave (MMW) exposure at 1.0 and 3.5 mW/cm2 on gap junction intercellular communication (GJIC) was studied in cultured HaCaT keratinocytes, using the fluorescence recovery after photobleaching (FRAP) technique and laser confocal scanning microscopy to follow the intracellular movement of 5,6-carboxyfluorescein diacetate dye. While MMW ... More
Characterization of phospholipase A2 activity in reticulocyte endocytic vesicles.
AuthorsBette-Bobillo P, Vidal M
JournalEur J Biochem
PubMed ID7883004
'Electron spin resonance spectroscopy was used to investigate the presence of phospholipase A2 activity in endocytic vesicles prepared from reticulocytes and to define some of its characteristics. Using spin-labeled phospholipid analogues, we measured the hydrolysis rate of the ester bond at position 2 during incubation with reticulocyte endocytic vesicles. We ... More
Direct measurement of acid permeation into rat oesophagus.
AuthorsTanaka S, Chu S, Hirokawa M, Montrose MH, Kaunitz JD
JournalGut
PubMed ID12740330
'BACKGROUND AND AIMS: The early responses of the oesophageal mucosa to acid perfusion may predict subsequent pathology. Mucosal responses to luminal acid may result either from acid permeating through the mucosa or from other unknown transduction mechanisms. In order to better understand the dynamics of acid permeation into the oesophageal ... More
Changes in cytoplasmic pH and in membrane potential in thrombin-stimulated human platelets.
AuthorsHorne WC, Norman NE, Schwartz DB, Simons ER
JournalEur J Biochem
PubMed ID7318826
'The response of human platelets to stimulation by a specific aggregant such as thrombin has been postulated to proceed sequentially via induction of response at the membrane, followed by execution of shape change, secretion, and aggregation of the platelets. We have shown earlier that the platelet response includes a depolarization ... More
Carboxyfluorescein diacetate labeling does not affect adhesion molecule expression or function in human neutrophils or eosinophils.
AuthorsDavenpeck KL, Chrest FJ, Sterbinsky SA, Bickel CA, Bochner BS
JournalJ Immunol Methods
PubMed ID8551041
'Fluorescently labeled leukocytes are commonly used in in vitro and in vivo experimental systems. However, the effects of fluorescent labeling on the expression and function of leukocyte adhesion molecules has not been examined in part because the extreme intensity of fluorescence tends to obscure signals from other fluorochromes used for ... More
Anomalous driving force for renal brush border H+/OH-transport characterized by using 6-carboxyfluorescein.
AuthorsVerkman AS, Ives HE
JournalBiochemistry
PubMed ID3013286
'The pH, delta pH, and membrane potential dependences of H+/OH-permeability in renal brush border membrane vesicles (BBMV) were studied by using the entrapped pH indicator 6-carboxyfluorescein (6CF). Quantitative H+/OH-fluxes (JH) were obtained from a calibration of the fluorescence response of 6CF to intravesicular pH using vesicles prepared with varying intravesicular ... More
Prediction of porcine semen fertility by homologous in vitro penetration (hIVP) assay.
AuthorsGadea J, Matás C, Lucas X
JournalAnim Reprod Sci
PubMed ID9877056
'A field trial was conducted to compare the fertility predicting capacity of different sperm assays applying classical semen analysis, sperm function and the homologous in vitro penetration test (hIVP) to 60 ejaculates from four boars collected over a period of 15 weeks. No differences were found between the groups of ... More
Lactic acid inhibition of gap junctional intercellular communication in in vitro astrocytes as measured by fluorescence recovery after laser photobleaching.
AuthorsAnders JJ
JournalGlia
PubMed ID2976397
'Lactic acid can permeate plasma membranes, causing intracellular acidosis. Gap junctions are sensitive to pHi and can be reversibly uncoupled by weak acids. In this study, dye coupling between in vitro astrocytes, presumably mediated by gap junctions, was measured in the absence and presence of lactic acid. Fluorescence recovery after ... More
Requirements for the Ca2+-independent component in the initial intercellular adhesion of C2 myoblasts.
AuthorsPizzey JA, Jones GE, Walsh FS
JournalJ Cell Biol
PubMed ID3198689
'Using a sensitive and quantitative adhesion assay, we have studied the initial stages of the intercellular adhesion of the C2 mouse myoblast line. After dissociation in low levels of trypsin in EDTA, C2 cells can rapidly reaggregate by Ca2+-independent mechanisms to form large multicellular aggregates. If cells are allowed to ... More
Novel fluorescein-based flow-cytometric method for detection of lipid peroxidation.
AuthorsMakrigiorgos GM, Kassis AI, Mahmood A, Bump EA, Savvides P
JournalFree Radic Biol Med
PubMed ID8958133
'The novel property of fluorescein to detect peroxyl radicals is demonstrated. On the basis of this observation, a fluorescein-based, flow-cytometric method to directly and continuously detect free radicals generated in cell membranes during lipid peroxidation has been developed. 5- and 6-Carboxyfluorescein (5-/6-CF) free in solution and fluorescein-labeled polylysine lose their ... More
Coupled Na+-H+ exchange in isolated acinar cells from rat exocrine pancreas.
AuthorsHellmessen W, Christian AL, Fasold H, Schulz I
JournalAm J Physiol
PubMed ID2990235
'Isolated acinar cells from the rat exocrine pancreas were loaded with 6-carboxyfluorescein diacetate (CFDA), and the intracellular pH (pHi) was estimated from the pH-dependent fluorescence intensity of trapped 6-carboxyfluorescein liberated from CFDA by intracellular esterases. The intracellular fluorescence intensity was calibrated by equilibrating the internal and external pH with nigericin ... More
Anti-HIV effects of chloroquine: inhibition of viral particle glycosylation and synergism with protease inhibitors.
AuthorsSavarino A, Lucia MB, Rastrelli E, Rutella S, Golotta C, Morra E, Tamburrini E, Perno CF, Boelaert JR, Sperber K, Cauda R
JournalJ Acquir Immune Defic Syndr
PubMed ID15076236
'OBJECTIVE: We tested the effects of chloroquine (CQ) on glycosylation of HIV particles and in combination with protease inhibitors (PIs) on HIV replication and on P-glycoprotein (P-gp)/multidrug resistance protein-1 (MRP1). DESIGN: CD4 cell lines were infected with laboratory strains and peripheral blood mononuclear cells were infected with primary isolates for ... More
Cell-penetrating peptides. A reevaluation of the mechanism of cellular uptake.
AuthorsRichard JP, Melikov K, Vives E, Ramos C, Verbeure B, Gait MJ, Chernomordik LV, Lebleu B
JournalJ Biol Chem
PubMed ID12411431
'Cellular uptake of a family of cationic cell-penetrating peptides (examples include Tat peptides and penetratin) have been ascribed in the literature to a mechanism that does not involve endocytosis. In this work we reevaluate the mechanisms of cellular uptake of Tat 48-60 and (Arg)(9). We demonstrate here that cell fixation, ... More
Application of gel microdroplet and flow cytometry techniques to selective enrichment of non-growing bacterial cells.
AuthorsManome A, Zhang H, Tani Y, Katsuragi T, Kurane R, Tsuchida T
JournalFEMS Microbiol Lett
PubMed ID11287142
'We describe an application of gel microdroplet (GMD) and flow cytometry techniques to selective enrichment of non-growing Leuconostoc mesenteroides cells, which are well culturable on other media, from a mixture with Bacillus subtilis cells in nutrient broth. After encapsulating cells of the mixed population within GMDs and a brief incubation ... More
Quantitative measurements of the efficacy of new anti-cancer agents on fresh human AML cells by using multivariate flow analysis.
AuthorsNakeff A, KuKuruga MA, Media JE, Valeriote F, Edelstein MB
JournalJ Exp Ther Oncol
PubMed ID9414426
'The testing of new human leukemia-specific drugs for activity against primary acute myeloid leukemia (AML) blasts is severely limited by the low and variable clonogenic potential of primary human leukemias in culture. To circumvent this problem, we have modified a previously described flow cytometric approach to permit the simultaneous determination ... More
Comparison of Flavobacterium and Sphingobacterium species by enzyme profiles, with use of pattern recognition of two-dimensional fluorescence data.
AuthorsPau CP, Patonay G, Moss CW, Hollis D, Carlone GM, Plikaytis BD, Warner IM
JournalClin Chem
PubMed ID3815801
'Enzyme profiles of eight Flavobacterium species and one Sphingobacterium species were compared after using a two-dimensional fluorescence technique. Enzyme contents and corresponding activities were rapidly determined for whole-cell preparations after incubation with a mixture of preselected fluorogenic substrates. A two-dimensional fluorescence spectrum of the resulting product mixture, measured with a ... More
A versatile flow cytometry-based assay for the determination of short- and long-term natural killer cell activity.
AuthorsJohann S, Blümel G, Lipp M, Förster R
JournalJ Immunol Methods
PubMed ID7561131
'A flow cytometry based method has been developed to assess natural killer (NK) cell activity in both short-term (4 h) and long-term (18 h) NK assays. Target cells were either labeled with PKH-2, c''FDA or D275. Simultaneously, dead cells were identified by counter-staining with the nuclear dye propidium iodide. Using ... More
A specific point mutant at position 1 of the influenza hemagglutinin fusion peptide displays a hemifusion phenotype.
AuthorsQiao H, Armstrong RT, Melikyan GB, Cohen FS, White JM
JournalMol Biol Cell
PubMed ID10436026
'We showed previously that substitution of the first residue of the influenza hemagglutinin (HA) fusion peptide Gly1 with Glu abolishes fusion activity. In the present study we asked whether this striking phenotype was due to the charge or side-chain volume of the substituted Glu. To do this we generated and ... More
Dynamic continuity of cytoplasmic and membrane compartments between plant cells.
AuthorsBaron-Epel O, Hernandez D, Jiang LW, Meiners S, Schindler M
JournalJ Cell Biol
PubMed ID3346323
'Fluorescence photobleaching was employed to examine the intercellular movement of fluorescein and carboxyfluorescein between contiguous soybean root cells (SB-1 cell line) growing in tissue culture. Results of these experiments demonstrated movement of these fluorescent probes between cytoplasmic (symplastic) compartments. This symplastic transport was inhibited with Ca2+ in the presence of ... More
A new sensitive and rapid automated fluorometric assay for detection of natural killer activity using carboxyfluorescein diacetate.
AuthorsSuzuki Y, Yoshikawa K, Yokochi T
JournalJ Immunoassay
PubMed ID2040709
'An automated fluorometric assay using carboxyfluorescein diacetate (CFDA) has been applied for the sensitive and rapid detection of natural killer (NK) activity. The lysis of target cells by NK cells was quantified by measuring the amount of CFDA released into the supernatant of culture wells with the aid of an ... More
Determination of infectious bursal disease virus titration and neutralization endpoints using fluorogenic staining.
AuthorsBayyari GR, Skeeles JK, Story JD, Slavik MF
JournalAvian Dis
PubMed ID1659366
'An automated method for determining infectious bursal disease virus (IBDV) titration and neutralization endpoints is described. The method employs the fluorogenic ester carboxyfluorescein-diacetate (CFDA) to stain cell monolayers in 96-well plates and a fluorescence-concentration analyzer. Titration results are compared with immunofluorescence and plaque assay titers. Virus-neutralization endpoint determination is objective, ... More
Evaluation of an automated method of percent reactive antibody determination.
AuthorsHeller MJ, Adams PW, Orosz CG
JournalHum Immunol
PubMed ID1293081
'A fluorescence-based automated method of percent reactive antibody (PRA) analysis is described. This method utilizes the conventional antibody-mediated, C''-dependent lymphocyte microcytotoxicity assay to detect alloantibodies, but replaces the eosin-based method for detection of cell death with a fluorescence-based method. To identify viable cells, lymphocytes were pretreated with carboxy fluorescein diacetate ... More
Pentamidine is active in vitro against Fusarium species.
AuthorsLionakis MS, Lewis RE, Samonis G, Kontoyiannis DP
JournalAntimicrob Agents Chemother
PubMed ID14506038
'Fusariosis is an emerging opportunistic mycosis against which currently used antifungals have limited activity. Here, we investigated the in vitro activities of pentamidine (PNT) against 10 clinical isolates of Fusarium species (five Fusarium solani isolates and five non-F. solani isolates) by using the National Committee for Clinical Laboratory Standards microdilution ... More
Flow cytometric comparison of the effects of trialkyltins on the murine erythroleukemic cell.
AuthorsZucker RM, Elstein KH, Easterling RE, Massaro EJ
JournalToxicology
PubMed ID2799821
'Cellular effects of exposure to tributyltin (TBT), triethyltin (TET), or trimethyltin (TMT) were investigated by flow cytometry employing the murine erythroleukemic cell (MELC) as a model cellular system. Cell viability was investigated by the carboxyfluorescein diacetate (CFDA) uptake/propidium iodide (PI) exclusion method: above a critical concentration (exposure for 4 h), ... More
Axon outgrowth is regulated by an intracellular purine-sensitive mechanism in retinal ganglion cells.
AuthorsBenowitz LI, Jing Y, Tabibiazar R, Jo SA, Petrausch B, Stuermer CA, Rosenberg PA, Irwin N
JournalJ Biol Chem
PubMed ID9792672
'Although purinergic compounds are widely involved in the intra- and intercellular communication of the nervous system, little is known of their involvement in the growth and regeneration of neuronal connections. In dissociated cultures, the addition of adenosine or guanosine in the low micromolar range induced goldfish retinal ganglion cells to ... More
Fusion of coated vesicles with lysosomes: measurement with a fluorescence assay.
AuthorsAltstiel L, Branton D
JournalCell
PubMed ID6131746
'A fluorescence assay was developed to measure the rate of fusion of highly purified clathrin-coated vesicles isolated from bovine brain with purified lysosomes isolated from bovine kidney. Coated vesicles and stripped vesicles, prepared by removal of clathrin from coated vesicles with dilute alkaline buffer, were labeled with the nonfluorescent dye ... More
Postnatal maturation of rabbit renal collecting duct: intercalated cell function.
AuthorsSatlin LM, Schwartz GJ
JournalAm J Physiol
PubMed ID3661716
'Intercalated cells play a major role in renal regulation of acid-base balance. We used fluorescent dyes to characterize postnatal maturation of intercalated cells. We stained rabbit collecting ducts with the pH-sensitive dye 6-carboxyfluorescein diacetate and identified individual intercalated cells by their bright green fluorescence. Number of fluorescent cells per millimeter ... More
Axial heterogeneity of intracellular pH in rat proximal convoluted tubule.
AuthorsPastoriza-Munoz E, Harrington RM, Graber ML
JournalJ Clin Invest
PubMed ID3036912
'In the proximal convoluted tubule (PT), the HCO3- reabsorptive rate is higher in early (EPS) compared with late proximal segments (LPS). To examine the mechanism of this HCO3- reabsorption profile, intracellular pH (pHi) was measured along the superficial PT of the rat under free-flow and stationary microperfusion using the pH-sensitive ... More
Fluorescent labeling of blood platelets in vivo.
AuthorsTangelder GJ, Slaaf DW, Reneman RS
JournalThromb Res
PubMed ID7167880
'In rabbits, a clear microscopic image of individual, fluorescent blood platelets flowing in a mesenteric microvessel could be obtained over the whole cross-sectional area of the vessel, following an intravenous injection of a solution containing 5-15 mg of the fluorochrome Acridine Red. Most of the circulating platelets were labeled. Activation ... More
Immortalized hypothalamic luteinizing hormone-releasing hormone neurons express a connexin 26-like protein and display functional gap junction coupling assayed by fluorescence recovery after photobleaching.
AuthorsMatesic DF, Germak JA, Dupont E, Madhukar BV
JournalNeuroendocrinology
PubMed ID8115017
'Expression of gap junction proteins was studied in the LHRH neuronal cell line, GT1-7, as a first step in defining the signalling mechanisms responsible for the pulsatile secretion of LHRH. GT1-7 cells were found to express a connexin 26-like protein that comigrated with mouse liver connexin 26 and that reacted ... More
Selective labeling of embryonic neurons cultured on astrocyte monolayers with 5(6)-carboxyfluorescein diacetate (CFDA).
AuthorsPetroski RE, Geller HM
JournalJ Neurosci Methods
PubMed ID8090014
'A method for selectively labeling cultured neurons using the vital dye, 5(6)-carboxyfluorescein diacetate (CFDA), is described. This non-fluorescent membrane-permeant dye is cleaved by cytosolic esterases into the fluorescent anion, 5(6)-carboxyfluorescein (CF). Both astrocytes and neurons exhibit brilliant fluorochromasia within minutes of CFDA loading. However, following a brief rinse in buffered ... More
Use of carboxyfluorescein diacetate to study formation of permeable channels between mouse blastomeres.
AuthorsGoodall H, Johnson MH
JournalNature
PubMed ID7057911
Targeting of small unilamellar liposomes to the galactose receptor in vivo.
AuthorsGregoriadis G, Senior J
JournalBiochem Soc Trans
PubMed ID6327432
Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ.
AuthorsThomas JA, Buchsbaum RN, Zimniak A, Racker E
JournalBiochemistry
PubMed ID36128
Fluorescence dye as monitor of internal pH in Escherichia coli cells.
AuthorsShechter E, Letellier L, Simons ER
JournalFEBS Lett
PubMed ID7042384
Carboxyfluorescein fluorochromasia assays. I. Non-radioactively labeled cell mediated lympholysis.
AuthorsBruning JW, Kardol MJ, Arentzen R
JournalJ Immunol Methods
PubMed ID6444653
Fluorescence microscopy methods for yeast.
AuthorsPringle JR, Preston RA, Adams AE, Stearns T, Drubin DG, Haarer BK, Jones EW
JournalMethods Cell Biol
PubMed ID2476649
Methods for studying the yeast vacuole.
AuthorsRoberts CJ, Raymond CK, Yamashiro CT, Stevens TH
JournalMethods Enzymol
PubMed ID1706462
Assessment of mitochondrial membrane potential in situ using single potentiometric dyes and a novel fluorescence resonance energy transfer technique.
AuthorsDykens JA, Stout AK
JournalMethods Cell Biol
PubMed ID11381600
Intercellular communication through gap junctions is reduced in senescent cells.
AuthorsXie HQ, Huang R, Hu VW
JournalBiophys J
PubMed ID1600097
Stimulus response coupling in human platelets: thrombin-induced changes in pHi.
AuthorsSimons ER, Schwartz DB, Norman NE,
JournalKroc Found Ser
PubMed ID6951954
Stimulus response coupling in human platelets: thrombin-induced changes in pHi.
Optimization of cytotoxic assay by target cell retention of the fluorescent dye carboxyfluorescein diacetate (CFDA) and comparison with conventional 51CR release assay.
AuthorsProvinciali M, Di Stefano G, Fabris N
JournalJ Immunol Methods
PubMed ID1401964
This paper describes the comparison between a fluorimetric NK assay based on the target cell retention of fluorescent dye carboxyfluorescein diacetate (cFDA) and standard 51Cr release assay. The results provide several suggestions to improve the cytotoxic assay based on the use of the fluorogenic substrate showing that the measurements of ... More
Development of an adhesive sheet for direct counting of bacteria on solid surfaces.
AuthorsYamaguchi N, Ishidoshiro A, Yoshida Y, Saika T, Senda S, Nasu M
JournalJ Microbiol Methods
PubMed ID12689718
An adhesive sheet was developed for direct counting of microorganisms on solid surfaces. The sheet consists of a polyurethane film base and water insoluble adhesive. SYBR Green II (for total direct counting) or 6-carboxyfluorescein diacetate (6CFDA) (for fluorescent vital staining) was used for fluorescent microscopy of bacteria collected on the ... More
Evaluation of changes in cytoplasmic pH in thrombin-stimulated human platelets.
AuthorsDavies TA, Dunn JM, Simons ER
JournalAnal Biochem
PubMed ID3434788
Thrombin causes a dose-dependent cytoplasmic alkalinization of normal human platelets. The pH probe 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) permits an easier and more accurate quantitation of the kinetics of this change previously measured with 9-aminoacridine and 6-carboxyfluorescein (6-CF). We report here a modification of the previously published 6-CF technique and confirm the thrombin-induced ... More
A simple fluorometric assay for quantifying the adhesion of tumour cells to endothelial monolayers.
AuthorsPrice EA, Coombe DR, Murray JC
JournalClin Exp Metastasis
PubMed ID7750203
A static adhesion assay employing 6-carboxy-3',6'-diacetylfluorescein (6-CFDA) as a fluorescent marker has been developed to study the interactions of tumour cell lines with endothelial monolayers. This assay allows simple, safe quantification of cell-cell adhesion using living cells. It has been used to demonstrate that the integrin adhesion molecule VLA-4 mediates ... More
A rapid, objective method for the detection of lymphocytotoxic antibodies using flow cytometry.
AuthorsTalbot D, Shenton BK, Givan AL, Proud G, Taylor RM
JournalJ Immunol Methods
PubMed ID3571990
Whilst several centres have reported lymphocytotoxic antibody detection using single and dual fluorescent stains with analysis of the fluorescence emitted from the cell population present in a well of a multiwell plate, problems are encountered with cell concentration and light emission overlap. A method we have developed using flow cytometry ... More
A fluorescence NK assay using flow cytometry.
AuthorsMcGinnes K, Chapman G, Marks R, Penny R
JournalJ Immunol Methods
PubMed ID3944470
A flow cytometric NK assay was developed in which the K562 targets were labelled with the fluorogenic substrate, carboxyfluorescein diacetate (c'FDA). This new assay compared favourably with results obtained using the conventional 51Cr-release assay. c'FDA was not toxic to target cells and did not inhibit lysis. The assay permits the ... More
Continuous monitoring of transport by fluorescence on cells and vesicles.
AuthorsEidelman O, Cabantchik ZI
JournalBiochim Biophys Acta
PubMed ID2686755
Fluorescence techniques are gaining wider applicability in the field of membrane transport due to their high temporal resolution, modest demand for biological material and the kinetic information which is made available by fluorescence tracings. The development of novel fluorescent substrates for particular transport systems and of novel fluorescent indicators for ... More
Fluorescent probes and flow cytometry: new insights into environmental bacteriology.
AuthorsPorter J, Deere D, Pickup R, Edwards C
JournalCytometry
PubMed ID8742166
Recent trends in flow cytometry have established new techniques in bacteriology. Advances in fluorescent dye technology complement these improvements, offering probes for a variety of cellular functions. Bacterial ecology requires the application of new techniques to help answer questions unanswerable by traditional methods alone. Here we review some aspects of ... More
Intracellular fluorescent labelling of cells for analysis of lymphocyte migration.
AuthorsBrenan M, Parish CR
JournalJ Immunol Methods
PubMed ID6438234
A procedure for analysing in vivo migration of lymphocytes labelled in vitro using intracellular fluorochromes is described. Comparison of carboxyfluorescein diacetate, a cytoplasmic label, with Hoechst dye No. 33342 (H33342), a DNA-binding fluorochrome, indicated that H33342 is superior. The concentration of H33342 used for labelling does not significantly affect viability ... More
Uptake of a fluorescent dye as a swift and simple indicator of organelle intactness: import-competent chloroplasts from soil-grown Arabidopsis.
AuthorsSchulz A, Knoetzel J, Scheller HV, Mant A
JournalJ Histochem Cytochem
PubMed ID15100247
We developed a rapid and reliable technique for specifically staining intact chloroplasts using the fluorescent dye carboxyfluorescein diacetate. Intact, import-competent chloroplasts were isolated simply and rapidly from soil-grown Arabidopsis thaliana plants, with yields of 20 +/- 5 micro g chlorophyll per g FW, greater than previously reported yields from soil-grown ... More
The heat shock-induced cell cycle arrest is attenuated by weak electromagnetic fields.
AuthorsTokalov SV, Gutzeit HO
JournalCell Prolif
PubMed ID12680877
Stress-induced effects in human acute leukaemia cells (HL-60) were studied by flow cytometry using the fluorescent dye carboxyfluorescein succinimidyl ester which allows the analysis of several successive cell generations for up to 10 days. Asynchronously cycling cells subjected to heat shock (30 min at 41 degrees C) responded in two ... More
Enzymatic quantification of cell-matrix and cell-cell adhesion.
AuthorsLöster K, Horstkorte R
JournalMicron
PubMed ID10568230
Adhesion assays are powerful tools to investigate the adhesive properties of cells. The quantification of cell adhesion enables determination of the capacity of cells to stick to a target, screening for novel adhesion involved binding molecules, exploration of structure-function relationships of adhesion molecules, evaluation of adhesion targets, and examination of ... More
Distribution of individual cytoplasmic pH values in a population of the yeast Saccharomyces cerevisiae.
AuthorsCimprich P, Slavík J, Kotyk A
JournalFEMS Microbiol Lett
PubMed ID7649447
Fluorescence ratio imaging microscopy using pH-sensitive fluorescent dyes makes it possible to evaluate statistical distribution of intracellular pH in a population of the yeast S. cerevisiae examined in a thin layer of suspension in a Petri dish. The distribution appears to fit a Gaussian curve with a half-width around the ... More
Identification of an E-selectin region critical for carbohydrate recognition and cell adhesion.
E-selectin elicits cell adhesion by binding to the cell surface carbohydrate, sialyl Lewis X (sLe(x)). We evaluated the effects of mutations in the E-selectin lectin domain on the binding of a panel of anti-E-selectin mAbs and on the recognition of immobilized sLe(x) glycolipid. Functional residues were then superimposed onto a ... More
Selection of bacteriophage-resistant mutants of Streptococcus thermophilus.
AuthorsViscardi M, Capparelli R, Di Matteo R, Carminati D, Giraffa G, Iannelli D
JournalJ Microbiol Methods
PubMed ID14500002
Phage-resistant mutants have been isolated from Streptococcus thermophilus. Selection was carried out using anti-phage antibodies or Hoechst 33258-labelled phages. Two mutants out of eight tested displayed reduced acidifying capacity. Selection of the bacteria that extruded more rapidly the fluorochrome 5-6-carboxyfluorescein diacetate (CFDA) restored the acidifying capacity of these two mutants ... More
Flow cytometric analysis of entire microbial colonies.
AuthorsSahar E, Nir R, Lamed R
JournalCytometry
PubMed ID8187581
Much could be gained if the scope of flow cytometry could be broadened to the study of entire cell colonies, rather than to populations of single, separate cells. This can be achieved by encapsulating single microbial cells in small spheres in a way that allows each cell to multiply and ... More
Improved microplate fluorometer counting of viable tumor and normal cells.
AuthorsRiordan HD, Riordan NH, Meng X, Zhong J, Jackson JA
JournalAnticancer Res
PubMed ID8074495
An improved method has been developed to count cells in situ based on the measurement of esterase activity with carboxyfluorescein diacetate. This sensitive, semiautomated microplate fluorometer assay was able to estimate viable cell numbers over a range of 5 x 10(2) to 2.6 x 10(5) cells/well in a tumor cell ... More
A rapid method for detection of cellular proliferation using carboxyfluorescein. Assay of growth factors (IL-2, IL-1) and growth inhibiting antibodies.
AuthorsHansson Y, Jacobson E, Ortlund J, Paulie S, Perlmann P
JournalJ Immunol Methods
PubMed ID3036952
The uptake of carboxyfluorescein diacetate (CFDA) into live cells was used as the basis for a simple, rapid and fully automated micromethod for determination of cell growth. The aim of the investigation was to adapt the CFDA method for detection of cell growth factors from cell culture supernatants. Thus, the ... More
A fluorescence photobleaching assay of gap junction-mediated communication between human cells.
AuthorsWade MH, Trosko JE, Schindler M
JournalScience
PubMed ID3961495
Gap junction-mediated communication between contiguous cells has been implicated in the regulation of cell proliferation and differentiation. This report describes a new technique to measure cell-cell communication, gap fluorescence redistribution after photobleaching, which is based on the diffusion-dependent return of 6-carboxyfluorescein-mediated fluorescence in a photobleached cell that is in contact ... More
Assessment of the morphology of frozen-thawed bull sperm in relation to its cryopreservation for artificial insemination.
AuthorsRevell S
JournalVet Rec
PubMed ID12846286
The morphology of sperm in raw semen was compared with that of the live sperm in semen which had been frozen and thawed. The thawed semen was stained with 6-carboxyfluorescein diacetate and propidium iodide and examined by fluorescence microscopy; smears of the raw semen were stained with eosin and nigrosin. ... More
Substituted benzyl acetates: a new class of compounds that reduce gap junctional conductance by cytoplasmic acidification.
AuthorsSpray DC, Nerbonne J, Campos de Carvalho A, Harris AL, Bennett MV
JournalJ Cell Biol
PubMed ID6736125
Conductance of gap junctions in many preparations has been shown to be sensitive to cytoplasmic pH, decreasing as pH decreases below 7.5 in fish and amphibian embryos and below 7.1 in crayfish septate axon. We have found a new class of compounds, benzyl acetate derivatives, that reversibly decrease junctional conductance, ... More
Comparison of different live/dead stainings for detection and quantification of adherent microorganisms in the initial oral biofilm.
AuthorsTawakoli PN, Al-Ahmad A, Hoth-Hannig W, Hannig M, Hannig C,
JournalClin Oral Investig
PubMed ID22821430
The aim of the present study was to investigate different fluorescence-based, two-color viability assays for visualization and quantification of initial bacterial adherence and to establish reliable alternatives to the ethidium bromide staining procedure. Bacterial colonization was attained in situ on bovine enamel slabs (n?=?6 subjects). Five different live/dead assays were ... More
Microengineered liver tissues for drug testing.
AuthorsKhetani SR, Berger DR, Ballinger KR, Davidson MD, Lin C, Ware BR,
Journal
PubMed ID25617027
Drug-induced liver injury (DILI) is a leading cause of drug attrition. Significant and well-documented differences between animals and humans in liver pathways now necessitate the use of human-relevant in vitro liver models for testing new chemical entities during preclinical drug development. Consequently, several human liver models with various levels of ... More
Application of glutaraldehyde for the staining of esterase-active cells with carboxyfluorescein diacetate.
AuthorsMorono Y, Takano S, Miyanaga K, Tanji Y, Unno H, Hori K
JournalBiotechnol Lett
PubMed ID15104134
Staining of esterase-active bacteria with carboxyfluorescein diacetate (CFDA) has been used to evaluate the viability of various types of cell. However, the outer membrane of Gram-negative bacteria prevents CFDA from permeating into the cell. Although EDTA can increase the permeability of the outer membrane allowing CFDA to enter the cells, ... More
An adhesion-associated agonist from the zona pellucida activates G protein-promoted elevations of internal Ca2+ and pH that mediate mammalian sperm acrosomal exocytosis.
AuthorsFlorman HM, Tombes RM, First NL, Babcock DF
JournalDev Biol
PubMed ID2504631
Solubilized oocyte zonae pellucidae promoted acrosomal exocytosis in fura-2- or carboxyfluorescein-loaded, mature bovine sperm. Associated elevations of internal [Ca2+] and pH in sperm suspensions were first detectable at 2-5 min, without apparent temporal resolution, and increased monotonically thereafter. Video imaging of fura-2-loaded, single cells identified a responsive subpopulation, destined to ... More
Calcium waves in astrocytes-filling in the gaps.
AuthorsFinkbeiner S
JournalNeuron
PubMed ID1351732
Stimulus-evoked cellular responses are sometimes organized in the form of propagating waves of cytoplasmic Ca2+ increase. Ca2+ waves can be elicited in cultured astrocytes by the neurotransmitter glutamate; however, the propagation mechanism is unknown. Here, qualitative and quantitative features of propagation suggest that astrocytic Ca2+ waves are mediated by an ... More
Detection of dead cells and measurement of cell killing by flow cytometry.
AuthorsKing MA
JournalJ Immunol Methods
PubMed ID10986413
A flow cytometer can quickly perform numerous quantitative, sensitive measurements on each individual cell within a large, heterogeneous population. The modern commercially available analytical instruments, which can be found in most hospitals, pathology laboratories, and cell biology research laboratories in the industrially developed countries, can now routinely measure fluorescence simultaneously ... More
Reduced cell-cell communication between mitotic and nonmitotic coupled cells.
AuthorsStein LS, Boonstra J, Burghardt RC
JournalExp Cell Res
PubMed ID1727042
The effects of mitosis on gap junctional intercellular communication (GJIC) were quantified in a clonal cell line of spontaneously immortalized rat granulosa cells (SIGC) using a fluorescence recovery after photobleaching assay. Reduction of GJIC was associated with the process of mitosis and was first apparent at the onset of prophase. ... More
A fluorescence microassay for the quantitation of integrin-mediated adhesion of neutrophil.
Authorsvan Kessel KP, Park CT, Wright SD
JournalJ Immunol Methods
PubMed ID8207264
The avidity of leukocyte integrin CR3 (Mac-1, CD11b/CD18, alpha m beta 2) on neutrophils (PMN) may be rapidly modulated by several agonists. We describe a method for determining the avidity of these receptors by measuring the adhesion of PMN to fibrinogen-coated surfaces. Cells are loaded with a succinimidyl ester of ... More
A new microcellular cytotoxicity test based on calcein AM release.
We present a microtest for cell-mediated immunity, based on the use of the Tarasaki tray and calcein AM vital dye. The number of target cells needed has been reduced to 500 per test with a corresponding tenfold reduction in the number of effector cells needed. Results were read at the ... More
Dieldrin inhibition of gap junctional intercellular communication in rat glial cells as measured by the fluorescence photobleaching and scrape loading/dye transfer assays.
AuthorsSuter S, Trosko JE, el-Fouly MH, Lockwood LR, Koestner A
JournalFundam Appl Toxicol
PubMed ID2446941
Application of the fluorescence-recovery after photobleaching (FRAP analysis) technique and scrape loading/dye transfer assay was made to measure the presence of gap junctional communication in primary rat glial cells in vitro in the presence and absence of the neurotoxicant and tumor promoter dieldrin, a chlorinated insecticide. Results demonstrate that primary ... More
Activation of sodium-proton exchange is a prerequisite for Ca2+ mobilization in human platelets.
AuthorsSiffert W, Akkerman JW
JournalNature
PubMed ID3027576
Stimulated platelets take up sodium ions and release hydrogen ions due to activation of Na+/H+ exchange resulting in cytoplasmic alkalinization. Suppression of Na+/H+ exchange either by removal of extracellular Na+ or by application of amiloride inhibits shape change, secretion of granule contents and aggregation. The data we present here indicate ... More
Intracellular turnover of fluorescein diacetate. Influence of membrane ionic gradients on fluorescein efflux.
AuthorsProsperi E
JournalHistochem J
PubMed ID2387757
The influence of the membrane ionic gradient on the efflux of Fluorescein after intracellular turnover of Fluorescein diacetate was studied in HeLa cells. The kinetics of Fluorescein efflux was monitored by determining with flow cytometry the decrease in fluorescence intensity of single cells. Alterations of the Na+ and K+ gradients ... More
Applications of automated simultaneous double fluorescence (SDF). II. HLA class I phenotyping using immunomagnetically separated T lymphocytes.
AuthorsLaundy GJ, Peberdy M, Klouda PT, Bradley BA
JournalJ Immunogenet
PubMed ID2687384
HLA class I phenotyping was performed using T-lymphocyte populations isolated by immunomagnetic beads (IMBs) coated with monoclonal antibodies with specificity for CD2, CD4 or CD8. The results were compared to those obtained using density gradient-separated lymphocytes (PBL). The typing trays were read by the automated simultaneous double-fluorescence (SDF) technique previously ... More
Nucleic acid detection using non-radioactive labelling methods.
AuthorsMansfield ES, Worley JM, McKenzie SE, Surrey S, Rappaport E, Fortina P
JournalMol Cell Probes
PubMed ID7477006
Nucleic acid probe-based assays are now widely used in genetic research, human identification, forensics and in a broad spectrum of clinical assays in the fields of microbiology, haematology/oncology and virology. Labelled probes are used in a variety of assay formats including dot-blots, Southern blots (DNA target), Northern blots (RNA target), ... More
The reversibility of tributyltin-induced toxicity in vitro as a function of concentration and duration of exposure (C x T).
AuthorsZucker RM, Massaro EJ, Elstein KH
JournalEnviron Res
PubMed ID1740093
The toxicity exhibited by murine erythroleukemic cells (MELC) exposed to tributyltin (TBT) is a function of both concentration (C) and duration of exposure (T). At or above a critical C x T product value (CPV) (e.g., 0.5-1.0 microM TBT, 6 hr), exposed MELC exhibit severe, irreversible toxicity: decreased membrane integrity ... More
Functional characterization of cell-to-cell coupling in cultured rat aortic smooth muscle.
AuthorsBlennerhassett MG, Kannan MS, Garfield RE
JournalAm J Physiol
PubMed ID3107400
Gap junction (GJ) occurrence and function was studied in cultured rat aortic smooth muscle cells, since cell-to-cell coupling is proposed to coordinate smooth muscle function but is difficult to study in the intact tissue. Cell proliferation in vitro formed a multilayered structure 10-15 cells thick. GJs connected cells to lateral ... More
A mitosis-specific phosphorylation of the gap junction protein connexin43 in human vascular cells: biochemical characterization and localization.
AuthorsXie H, Laird DW, Chang TH, Hu VW
JournalJ Cell Biol
PubMed ID9105048
Western blotting studies revealed that connexin43 (Cx43), one of the major gap junction proteins in human vascular endothelial cells, is posttranslationally modified during mitosis. This mitosis-specific modification results in a Cx43 species that migrates as a single protein band and was designated Cx43(m). Cx43(m) was shown to be the result ... More
MRP2, a human conjugate export pump, is present and transports fluo 3 into apical vacuoles of Hep G2 cells.
AuthorsCantz T, Nies AT, Brom M, Hofmann AF, Keppler D
JournalAm J Physiol Gastrointest Liver Physiol
PubMed ID10762605
The multidrug resistance protein 2 (MRP2, symbol ABCC2) transports anionic conjugates and certain amphiphilic anions across the apical membrane of polarized cells. Human hepatoma Hep G2 cells retain hepatic polarity and form apical vacuoles into which cholephilic substances are secreted. Immunofluorescence microscopy showed that human MRP2 was expressed in the ... More
Generic liposome reagent for immunoassays.
AuthorsPlant AL, Brizgys MV, Locasio-Brown L, Durst RA
JournalAnal Biochem
PubMed ID2742132
We have derivatized liposomes with antibodies by using avidin to crosslink biotinylated phospholipid molecules in the liposome membranes with biotinylated antibody molecules. A comparison of the biotin binding activity of avidin in solution and avidin associated with liposomes shows that avidin bound to biotinylated phospholipid in liposome membranes retains full ... More
Novel fluorescent broth microdilution method for fluconazole susceptibility testing of Candida albicans.
AuthorsLiao RS, Rennie RP, Talbot JA
JournalJ Clin Microbiol
PubMed ID11427602
A comparative evaluation of the reference National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution method with a novel fluorescent carboxyfluorescein diacetate (CFDA)-modified microdilution method for the susceptibility testing of fluconazole was conducted with 68 Candida strains, including 53 Candida albicans, 5 Candida tropicalis, 5 Candida glabrata, and 5 Candida ... More
Intracellular pH of the turtle bladder assessed with fluorescent probes.
AuthorsArruda JA, Wheeler RP, Dytko G, Talor Z
JournalMiner Electrolyte Metab
PubMed ID3696088
Intracellular pH of the turtle bladder was measured with fluorescent probe 6-carboxyfluorescein (6-CF) diacetate. In isolated cells this probe provides reliable, reproducible and fast measurements of intracellular pH. The probe was mainly located in the cytosol and thus the values of intracellular pH mainly reflect cytosolic pH. The values of ... More
Concentration/response effect of 2,2', 4,4', 5,5'-hexabromobiphenyl on cell-cell communication in vitro: assessment by fluorescence redistribution after photobleaching ("FRAP").
AuthorsEvans MG, Trosko JE
JournalCell Biol Toxicol
PubMed ID2852992
Inhibition of gap junction-mediated cell-cell communication might be a mechanism for several types of cellular dysfunctions, including tumor promotion. Although many different assays have been designed to measure gap junction-mediated intercellular communication, we applied a new technique, termed Fluorescence Redistribution After Photobleaching ("FRAP"), to assess the ability of a known ... More